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| Product Name | Nicking Endonuclease Nt.BbvCI (Recognition Site: 5'-CCTCAGC-3', Top-Strand Nicking) |
| Catalog No. | IVDRM-0164 |
| Description | Nt.BbvCI is a nicking endonuclease that recognizes the sequence 5'-CCTCAGC-3' and cleaves only one strand (the top strand) of a double-stranded DNA substrate, introducing a single-stranded nick rather than cutting the duplex through. The enzyme is widely used in isothermal nucleic acid amplification methods such as SDA and RCA: the nick generated by Nt.BbvCI primes polymerase-mediated strand displacement, and repeated cycles of nicking, displacement, and extension drive exponential amplification of the target nucleic acid. |
| Intended Use | For molecular cloning and nucleic acid isothermal amplification applications (e.g., SDA, RCA), where enzyme-generated nicks trigger polymerase strand displacement to achieve exponential amplification. |
| Principle / Technology | Nicking endonuclease: cleaves only one strand of the dsDNA substrate. Nt.BbvCI nicks the top strand within its recognition site (5'-CC↓TCAGC-3'), producing a nick rather than a double-strand break. |
| Sample Type | Double-stranded DNA (dsDNA) substrates. |
| Performance Range / Specifications | Concentration: 5 U/µL. One unit (U) is the amount of enzyme required to completely convert 1 µg of supercoiled p615 DNA to the open circular form in 1 hour at 37°C in a 50 µL reaction. |
| Specificity | Recognizes 5'-CCTCAGC-3' and nicks the top strand. |
| Reaction Conditions / Protocol | 1× reaction buffer; incubate at 37°C. Heat inactivation: 80°C for 20 min. |
| Components / Formulation | Nt.BbvCI nicking endonuclease, 5 U/µL; supplied with the recommended reaction buffer. |
| Storage Conditions | Store at -25°C to -15°C. |
| Shelf Life | 2 years when stored at -25°C to -15°C. |
| Package Specifications | 150 U |
| Concentration | 5 U/µL |
| Activity / Unit Definition | One unit (U) is defined as the amount of enzyme required to completely convert 1 µg of supercoiled p615 DNA into the open circular form within 1 hour at 37°C in a 50 µL reaction. |
| Expiration Date / Stability | Stable for 2 years at -25°C to -15°C. |
| Recommended Buffer System | 1× universal restriction enzyme reaction buffer (supplied). |
| Application Notes / Precautions | Incubate reactions at 37°C for optimal activity. Inactivate the enzyme at 80°C for 20 min before downstream steps. Avoid repeated freeze-thaw cycles; store at -25°C to -15°C. |
| Recognition Site | 5'-CC↓TCAGC-3' 3'-GG AGTCG-5' (Nt.BbvCI cleaves the top strand, as indicated by ↓.) |
| Heat Inactivation | 80°C for 20 min. |
For research use only, not for clinical use.
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